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Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis

Schizosaccharomyces pombe, also known as fission yeast, is an established model for studying chromosome biological processes. Over the years, research employing fission yeast has made important contributions to our knowledge about chromosome segregation during meiosis, as well as meiotic recombinati...

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Autores principales: Li, Dmitriy, Roca, Marianne, Yuecel, Raif, Lorenz, Alexander
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6823302/
https://www.ncbi.nlm.nih.gov/pubmed/30739171
http://dx.doi.org/10.1007/s00412-019-00691-y
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author Li, Dmitriy
Roca, Marianne
Yuecel, Raif
Lorenz, Alexander
author_facet Li, Dmitriy
Roca, Marianne
Yuecel, Raif
Lorenz, Alexander
author_sort Li, Dmitriy
collection PubMed
description Schizosaccharomyces pombe, also known as fission yeast, is an established model for studying chromosome biological processes. Over the years, research employing fission yeast has made important contributions to our knowledge about chromosome segregation during meiosis, as well as meiotic recombination and its regulation. Quantification of meiotic recombination frequency is not a straightforward undertaking, either requiring viable progeny for a genetic plating assay, or relying on laborious Southern blot analysis of recombination intermediates. Neither of these methods lends itself to high-throughput screens to identify novel meiotic factors. Here, we establish visual assays novel to Sz. pombe for characterizing chromosome segregation and meiotic recombination phenotypes. Genes expressing red, yellow, and/or cyan fluorophores from spore-autonomous promoters have been integrated into the fission yeast genomes, either close to the centromere of chromosome 1 to monitor chromosome segregation, or on the arm of chromosome 3 to form a genetic interval at which recombination frequency can be determined. The visual recombination assay allows straightforward and immediate assessment of the genetic outcome of a single meiosis by epi-fluorescence microscopy without requiring tetrad dissection. We also demonstrate that the recombination frequency analysis can be automatized by utilizing imaging flow cytometry to enable high-throughput screens. These assays have several advantages over traditional methods for analyzing meiotic phenotypes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00412-019-00691-y) contains supplementary material, which is available to authorized users.
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spelling pubmed-68233022019-11-06 Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis Li, Dmitriy Roca, Marianne Yuecel, Raif Lorenz, Alexander Chromosoma Original Article Schizosaccharomyces pombe, also known as fission yeast, is an established model for studying chromosome biological processes. Over the years, research employing fission yeast has made important contributions to our knowledge about chromosome segregation during meiosis, as well as meiotic recombination and its regulation. Quantification of meiotic recombination frequency is not a straightforward undertaking, either requiring viable progeny for a genetic plating assay, or relying on laborious Southern blot analysis of recombination intermediates. Neither of these methods lends itself to high-throughput screens to identify novel meiotic factors. Here, we establish visual assays novel to Sz. pombe for characterizing chromosome segregation and meiotic recombination phenotypes. Genes expressing red, yellow, and/or cyan fluorophores from spore-autonomous promoters have been integrated into the fission yeast genomes, either close to the centromere of chromosome 1 to monitor chromosome segregation, or on the arm of chromosome 3 to form a genetic interval at which recombination frequency can be determined. The visual recombination assay allows straightforward and immediate assessment of the genetic outcome of a single meiosis by epi-fluorescence microscopy without requiring tetrad dissection. We also demonstrate that the recombination frequency analysis can be automatized by utilizing imaging flow cytometry to enable high-throughput screens. These assays have several advantages over traditional methods for analyzing meiotic phenotypes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00412-019-00691-y) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2019-02-09 2019 /pmc/articles/PMC6823302/ /pubmed/30739171 http://dx.doi.org/10.1007/s00412-019-00691-y Text en © The Author(s) 2019 OpenAccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Article
Li, Dmitriy
Roca, Marianne
Yuecel, Raif
Lorenz, Alexander
Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis
title Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis
title_full Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis
title_fullStr Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis
title_full_unstemmed Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis
title_short Immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis
title_sort immediate visualization of recombination events and chromosome segregation defects in fission yeast meiosis
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6823302/
https://www.ncbi.nlm.nih.gov/pubmed/30739171
http://dx.doi.org/10.1007/s00412-019-00691-y
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